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PubMed2005

Salivary Actinomyces naeslundii genospecies 2 and Lactobacillus casei levels predict pregnancy outcomes.

BACKGROUND: Gravida's poor periodontal health is emerging as a modifiable independent risk factor for preterm delivery and low birth weight. METHODS: To test the hypothesis that oral bacteria other than periodontal pathogens are also associated with pregnancy outcomes, specific oral bacterial levels measured during pregnancy were evaluated in relation to gestational age and birth weight while controlling for demographic, medical, and dental variables. The study population consisted of 297 predominantly African- American women who were pregnant for the first time. The salivary bacterial levels evaluated were Streptococcus mutans, Streptococcus sobrinus, Streptococcus sanguinus, Lactobacillus acidophilus, Lactobacillus casei, Actinomyces naeslundii genospecies (gsp) 1 and 2, total streptococci, and total cultivable organisms. RESULTS: For 1 unit increase in log(10) A. naeslundii gsp 2 levels, there was a 60 gm decrease in birth weight (beta = -59.7 g; SE = 29.1; P = 0.04), and a 0.17 week decrease in gestational age (beta = -0.17 wk; SE = 0.09; P = 0.05). In contrast, per 1 unit increase in log(10) L. casei levels, there was a 42 gm increase in birth weight (beta = 42.2 g; SE = 19.3; P = 0.03), and a 0.13 week increase in gestational age (beta = 0.13 week; SE = 0.06; P = 0.04). CONCLUSIONS: We conclude that other oral bacterial species can also be related to pregnancy outcomes in addition to previously reported periodontal pathogens. These organism levels may not only predict poor pregnancy outcomes, but also be used as modifiable risk factors in reducing prematurity and low birth weight.

Actinomyces↗

PubMedDate not supplied

Mutans streptococcal serotypes in children with gastroesophageal reflux disease.

It has been suggested that vomiting acid gastric contents in bulimia might favor oral growth of Streptococcus sobrinus. We studied the colonization of Streptococcus sobrinus (serotypes g and d) and Streptococcus mutans (serotypes c, e and f) in sixteen children, ages five to fifteen years, who had suffered for four to eleven years from gastroesophageal reflux, another condition with recurrent acid regurgitation. Our aim was to find out if the prevalence of Streptococcus sobrinus would be higher also in this patient group. Mutants streptococci were detected in twelve out of sixteen (75 percent) study patients of the saliva samples cultured on MSB agar. For the Mutans streptococci positive children healthy controls were matched by salivary levels of mutans streptococci and age as closely as possible. From each child three to six isolates representing both Streptococcus mutans and Streptococcus sobrinus (n = 103) were serotyped by immunodiffusion method. The distribution of serotypes in the study/control group was: c: 7/10; e: 4/2; f: 0/1; g:3/2; d:0/0. One strain in the study group remained untypable. All patients infected with Streptococcus sobrinus were also infected with Streptococcus mutans. Our results indicate the great similarity in the distribution of ms serotypes in the gastroesophageal reflux children and their healthy controls. The data do not suggest that the acid regurgitation would have an influence on the prevalence of Streptococcus sobrinus.

Adolescent↗

PubMedDate not supplied

Effects of phenytoin on dental caries and plaque in Streptococcus sobrinus-infected rats.

Effects of phenytoin (PHT) administration on dental caries in rats infected with Streptococcus sobrinus 6715 were investigated. Twenty-day-old specific pathogen-free Fischer male rats were infected with S. sobrinus 6715 and fed diet 2000 containing 56% sucrose with or without PHT for 52 days. Antibody responses against anti-S. sobrinus in serum and saliva failed to show a statistical difference between PHT-treated and nontreated rats. These results indicate that PHT treatment increased plaque deposition and dental caries in the rats infected with S. sobrinus 6715 and fed diet 2000 containing PHT (1-2 mg/g), as compared with those similarly infected and fed diet without PHT (P < 0.001).

Animals↗

PubMed1995

Intrafamilial similarity in immunoblot profiles of salivary immunoglobulin A antibody activity to oral streptococci.

Analysis of the salivary immunoglobulin A (IgA) antibody activity to antigens separated from oral streptococci was carried out on 49 individuals in 11 families. The aim of this study were to i) study the human salivary IgA activity within families to antigens separated from reference strains of Streptococcus mutans, Streptococcus sobrinus and Streptococcus parasanguis and ii) to search for potential differences in the salivary IgA activity to the streptococcal antigens several years apart. The immunoblot revealed similarity in saliva IgA response within the families. A majority of the bands, approximately 7 (median) for each extract was found in both child and parent. A few bands (approximately 3) were found in parents but not in children. Only one to two bands were found in children's saliva without a corresponding band in any of the parents. The antibody activity of saliva samples obtained several years apart was essentially unchanged, especially considering the total number of bands. Differences in the relative intensity of the bands could be seen, and a few bands appeared only in some immunoblots for a specific subject. For 3 of 4 children participating in the longitudinal study, more bands against mutans streptococci appeared with age.

Adolescent↗

PubMed1992

Effects of food preservatives on growth and metabolism of plaque bacteria in vitro and in vivo.

The aims of the present study were to assess the consumption of food preservatives during the last decades, and to study the effect of the preservatives, sorbic and benzoic acid, on growth and glycolysis of oral bacteria in vitro, and on acid formation by dental plaque in vivo. Five consumption reports from the Central Bureau of Statistics of Norway were used to estimate alterations in consumption of staple food containing the two preservatives. A modified broth dilution method was used to determine the MIC values of the preservatives against Streptococcus sobrinus and Streptococcus sanguis. Extracellular 14C-glycolytic metabolites were studied by HPLC analyses. Plaque-pH measurements were used to assess possible effects on acid production. The consumption reports indicated increased consumption of preservatives. The in vitro testing suggested that legal concentrations of preservatives may inhibit the growth of oral streptococci. However, the preservatives did not inhibit in vitro glycolysis at tested concentrations. In vivo testing with similar concentrations (0.4% w/v) showed a significant effect. A higher concentration (2% w/v potassium sorbate) had a tendency to inhibit acid-formation by dental plaque even more.

Acids↗

PubMed2001

Spatial arrangements and associative behavior of species in an in vitro oral biofilm model.

The spatial arrangements and associative behavior of Actinomyces naeslundii, Veillonella dispar, Fusobacterium nucleatum, Streptococcus sobrinus, and Streptococcus oralis strains in an in vitro model of supragingival plaque were determined. Using species-specific fluorescence-labeled antibodies in conjunction with confocal laser scanning microscopy, the volumes and distribution of the five strains were assessed during biofilm formation. The volume-derived cell numbers of each strain correlated well with respective culture data. Between 15 min and 64 h, populations of each strain increased in a manner reminiscent of batch growth. The microcolony morphologies of all members of the consortium and their distributions within the biofilm were characterized, as were interspecies associations. Biofilms formed 15 min after inoculation consisted principally of single nonaggregated cells. All five strains adhered strongly to the saliva-conditioned substratum, and therefore, coadhesion played no role during the initial phase of biofilm formation. This observation does not reflect the results of in vitro coaggregation of the five strains, which depended upon the nature of the suspension medium. While the possibility cannot be excluded that some interspecies associations observed at later stages of biofilm formation were initiated by coadhesion, increase in bacterial numbers appeared to be largely a growth phenomenon regulated by the prevailing cultivation conditions.

Actinomyces↗

PubMed2001

Validation of an in vitro biofilm model of supragingival plaque.

The study of biofilm structure and function mandates the use of model systems for which a host of environmental variables can be rigorously controlled. We describe a model of supragingival plaque containing Actinomyces naeslundii, Veillonella dispar, Fusobacterium nucleatum, Streptococcus sobrinus, and Streptococcus oralis wherein cells are cultivated anaerobically in a saliva-based medium on hydroxyapatite discs coated with a salivary pellicle, with material and pieces of apparatus common to all microbiology laboratories. After 0.5 hr, 16.5 hrs, 40.5 hrs, and 64.5 hrs, the composition of adherent biofilms was analyzed by culture techniques, live/dead fluorescence staining, and confocal laser scanning microscopy. Repeated independent trials demonstrated the repeatability of biofilm formation after 40.5 hrs and 64.5 hrs. Brief exposures of biofilms to chlorhexidine or Triclosan produced losses in viability similar to those observed in vivo. This biofilm model should prove very useful for pre-clinical testing of prospective anti-plaque agents at clinically relevant concentrations.

Actinomyces↗

PubMed2004

Influence of resin monomers on growth of oral streptococci.

Ethyleneglycol dimethacrylate monomers have been previously reported to stimulate the growth of certain caries-associated bacteria on the basis of turbidity measurements. To elucidate the detail of this effect, we examined the influence of resin monomers on the growth of Streptococcus sobrinus and Streptococcus sanguis by determination of bacterial numbers (colony-forming units), morphological observation, and chemical analysis. Although the absorbance values in the stationary phase of bacterial suspension were increased in the presence of ethyleneglycol monomers, no significant differences were observed for bacterial numbers throughout the incubation period. Scanning electron microscopy observation revealed the formation of sparse vesicular material surrounding bacterial cells when incubated with ethyleneglycol monomers, and these products were proved to be resin polymers. The results demonstrate that the apparent biomass increase during incubation with ethyleneglycol monomers is due not to promotion of bacterial multiplication, but to the polymerization of resin monomers to form vesicular structures attached to cells.

Acrylates↗

PubMed1991

Effects of propolis on dental caries in rats.

Propolis, the resinous hive product collected by bees, is important in the defense of the hive. The effects of propolis on growth and glucosyltransferase activity of Streptococcus sobrinus 6715, Streptococcus mutans PS14 and Streptococcus cricetus OMZ61 in vitro, and on dental caries in rats infected with S. sobrinus 6715 were investigated. Propolis had antimicrobial activity against S. sobrinus, S. mutans and S. cricetus, and inhibited both water-insoluble glucan synthesis and glucosyltransferase activity. In rats inoculated with S. sobrinus, about half of their fissures were carious, while dental caries was significantly less (p = 0.01) in rats given propolis. Dental caries was markedly decreased by the multiple actions of propolis which had antimicrobial activity, inhibited water-insoluble glucan synthesis, and inhibited glucosyltransferase activity. No toxic effects of propolis on the growth of rats were observed under experimental conditions in this study. These results suggest that propolis can control dental caries in the rat model system.

Animals↗

PubMed2002

Real-time PCR for quantification of Streptococcus mutans.

A real-time polymerase chain reaction (PCR) assay was developed for the quantification of Streptococcus mutans. Primers targeting gtf genes of S. mutans were designed and tested for their specificity using 28 oral streptococcal strains, three other bacterial strains, and human DNA. The primers could amplify specifically the target DNA fragment from a mixture of oral streptococcus genomic DNA containing about 10 fg to 10 ng of S. mutans genome DNA. The real-time PCR produced a linear quantitative detection range over concentrations spanning seven exponential values, with a detection limit of a few copies of S. mutans' genomic DNA per reaction tube. The results of the real-time PCR assay corresponded well to those of conventional culture assays for S. mutans in saliva samples. A real-time PCR assay for Streptococcus sobrinus and Streptococcus downei was also established and produced results that corresponded well to those from conventional culture assays for S. sobrinus in saliva samples. These assays will be useful as a new means to assess one of the important risk factors for caries.

Bacterial Typing Techniques↗

PubMed1996

Inhibitory effects of funoran on the adherence and colonization of mutans streptococci.

Funoran, a sulfated polysaccharide extracted from the seaweed Gloiopeltis furcata strongly inhibited the absorption of mutans streptococci to saliva-coated hydroxyapatite (S-HA), but enhanced that of Streptococcus sanguis ATCC 10556 and Streptococcus oralis ATCC 10557. Furthermore, funoran had a strong desorption activity against mutans streptococci preadsorbed to S-HA. In the presence of sucrose, the absorption of cells of Streptococcus sobrinus B13, Streptococcus mutans Ingbritt, and S. mutans MT8148R to S-HA was strongly inhibited by 0.01% funoran. The colonization of S. sobrinus 6715 inoculated on the molar teeth of experimental rats that were administered funoran was less frequent than that in a funoran-free group. The mean buccal and lingual, sulcal, and total caries scores of rat groups administered funoran were significantly lower than those of the funoran-free group.

Adsorption↗

PubMed1994

Immunological characteristics of a synthetic peptide associated with a catalytic domain of mutans streptococcal glucosyltransferase.

The immunogenicity of a multiple antigenic peptide construct consisting of four copies of the synthetic 21-mer peptide DANFDSIRVDAVDNVDADLLQ was measured. The composition of this peptide was derived from a sequence in the N-terminal region of mutans streptococcal glucosyltransferases (GTFs) containing an aspartic acid implicated in catalysis. The peptide (CAT) construct was synthesized as a tetramer on a lysine backbone and subcutaneously injected into Sprague-Dawley rats for polyclonal antibody formation or intraperitoneally injected into BALB/c mice, and then spleen cell fused with Sp2/0Ag14 murine myeloma cells for monoclonal antibody formation. The resulting rat antisera and mouse monoclonal antibodies reacted with CAT and with native GTF isozymes from Streptococcus sobrinus and Streptococcus mutans (in enzyme-linked immunosorbent assay and Western blot [immunoblot] analyses). Functional inhibition of the water-insoluble glucan synthetic activity of S. sobrinus GTF-I was demonstrated with an immunoglobulin M anti-CAT monoclonal antibody (> 80% inhibited) and with rat sera (approximately 17% inhibited). The monoclonal antibody preparation also modestly inhibited the water-soluble glucan synthetic activity of an S. mutans GTF mixture. These results suggest that the CAT peptide contains B-cell epitopes that are similar to those of intact mutans streptococcal GTFs and has the potential to elicit antibody that can inhibit GTF function. Thus, sequences within this peptide construct may have value for inclusion in a synthetic dental caries vaccine.

Amino Acid Sequence↗

PubMed2006

Effect of mixed mutans streptococci colonization on caries development.

OBJECTIVE: To evaluate the clinical importance of mixed mutans streptococci colonization in predicting caries in preschool children. METHODS: Caries prevalence was examined twice, with a 6-month interval, in 410 preschool children aged 3-4 years at baseline. A commercial strip method was used to evaluate the mutans streptococci score in plaque collected from eight selected interdental spaces and in saliva. Mutans streptococci typing polymerase chain reaction (PCR) assays (Streptococcus sobrinus and Streptococcus mutans, including serotypes c, e, and f) were performed using colonies on the strips as template. RESULTS: Twenty variables were examined in a univariate analysis to predict caries development: questionnaire variables, results of clinical examination, mutans streptococci scores, and PCR detection of S. sobrinus and S. mutans (including serotypes c, e, and f). Sixteen variables showed statistically significant associations (P < 0.04) in the univariate analysis. However, when entered into a logistic regression, only five variables remained significant (P < 0.05): caries experience at baseline; mixed colonization of S. sobrinus and S. mutans including S. mutans serotypes; high plaque mutans streptococci score; habitual use of sweet drinks; and nonuse of fluoride toothpaste. CONCLUSION: 'Mixed mutans streptococci colonization' is a novel measure correlated with caries development in their primary dentition.

Beverages↗

PubMed2005

Low salivary IgA activity to cell-surface antigens of mutans streptococci related to HLA-DRB1*04.

BACKGROUND/AIMS: Mutans streptococci are found in almost all individuals, though there are large differences in colonization levels between individuals. These differences are not readily explained, though several factors are believed to influence the colonization. One factor is the immune response to mutans streptococci, mainly provided by salivary immunoglobulin A (IgA). In a previous study, differences in salivary IgA reactions to oral streptococci were observed between human leukocyte antigen (HLA)-DR4-positive and DR4-negative individuals. A lower salivary IgA activity to Streptococcus mutans in particular was most pronounced for two DR4 subgroups, DRB1*0401 and *0404. The main purpose of this study was to further investigate, in a larger study group, the salivary IgA activity to antigens of three oral streptococci in relation to different HLA-DRB1*04 alleles. METHODS: Stimulated saliva was collected from 58 HLA-DRB1*04-positive individuals. Whole cell antigen extracts from S. mutans, Streptococcus sobrinus and Streptococcus parasanguis and the streptococcal antigen (SA) I/II were separated in SDS-PAGE, transblotted and detected with diluted saliva (Western blot), and analyzed in a computer program. All distinct immunoblot bands over 100 kDa were recorded and compared in relation to DRB1*04. RESULTS: The immunoblots revealed lower salivary IgA reactions to S. mutans, S. sobrinus and SA I/II, but not to S. parasanguis, for the DRB1*0401- and *0404-positive individuals compared to other DRB1*04 types. For the *0401 subgroup there was a significant association with a lower IgA response to S. mutans. CONCLUSION: The results confirm earlier observations and may also support previous demonstrated association between colonization by mutans streptococci and the serologically defined HLA-DR4.

Adult↗

PubMed2001

Analysis of bacterial detachment from substratum surfaces by the passage of air-liquid interfaces.

A theoretical analysis of the detachment of bacteria adhering to substratum surfaces upon the passage of an air-liquid interface is given, together with experimental results for bacterial detachment in the absence and presence of a conditioning film on different substratum surfaces. Bacteria (Streptococcus sobrinus HG1025, Streptococcus oralis J22, Actinomyces naeslundii T14V-J1, Bacteroides fragilis 793E, and Pseudomonas aeruginosa 974K) were first allowed to adhere to hydrophilic glass and hydrophobic dimethyldichlorosilane (DDS)-coated glass in a parallel-plate flow chamber until a density of 4 x 10(6) cells cm(-2) was reached. For S. sobrinus HG1025, S. oralis J22, and A. naeslundii T14V-J1, the conditioning film consisted of adsorbed salivary components, while for B. fragilis 793E and P. aeruginosa 974K, the film consisted of adsorbed human plasma components. Subsequently, air bubbles were passed through the flow chamber and the bacterial detachment percentages were measured. For some experimental conditions, like with P. aeruginosa 974K adhering to DDS-coated glass and an air bubble moving at high velocity (i.e., 13.6 mm s(-1)), no bacteria detached upon passage of an air-liquid interface, while for others, detachment percentages between 80 and 90% were observed. The detachment percentage increased when the velocity of the passing air bubble decreased, regardless of the bacterial strain and substratum surface hydrophobicity involved. However, the variation in percentages of detachment by a passing air bubble depended greatly upon the strain and substratum surface involved. At low air bubble velocities the hydrophobicity of the substratum had no influence on the detachment, but at high air bubble velocities all bacterial strains were more efficiently detached from hydrophilic glass substrata. Furthermore, the presence of a conditioning film could either inhibit or stimulate detachment. The shape of the bacterial cell played a major role in detachment at high air bubble velocities, and spherical strains (i.e., streptococci) detached more efficiently than rod-shaped organisms. The present results demonstrate that methodologies to study bacterial adhesion which include contact with a moving air-liquid interface (i.e., rinsing and dipping) yield detachment of an unpredictable number of adhering microorganisms. Hence, results of studies based on such methodologies should be referred as "bacterial retention" rather than "bacterial adhesion".

Actinomyces↗

PubMed1993

Inhibitory effect of oolong tea polyphenols on glycosyltransferases of mutans Streptococci.

Oolong tea extract (OTE) was found to inhibit the water-insoluble glucan-synthesizing enzyme, glucosyltransferase I (GTase-I), of Streptococcus sobrinus 6715. The GTase-inhibitory substance in the OTE was purified successive adsorption chromatography on Diaion HP-21 and HP-20 columns; this was followed by further purification by Sephadex LH-20 column chromatography. A major fraction that inhibited GTase activity (fraction OTF10) was obtained, and the chemical analysis of OTF10 indicated that it was a novel polymeric polyphenol compound that had a molecular weight of approximately 2,000 and differed from other tea polyphenols. Catechins and all other low-molecular-weight polyphenols except theaflavin derived from balck tea did not show significant GTase-inhibitory activities. It was found that OTE amd PTF10 markedly inhibit GTase-I and yeast alpha-glucosidase, but not salivary alpha-amylase. Various GTases purified from S. sobrinus and Streptococcus mutans were examined for inhibition by OTE and OTF10. It was determined that S. sobrinus GTase-I and S. mutans cell-free GTase synthesizing water-soluble glucan were most susceptible to the inhibitory action of OTF10, while S. sobrinus GTase-Sa and S. mutans cell-associated GTase were moderately inhibited; no inhibition of S. sobrinus GTase-Sb was observed. Inhibition of a specific GTase or specific GTases of mutants streptococci resulted in decreased adherence of the growing cells of these organisms. The inhibitory effect of OTF10 on cellular adherence was significantly stronger than that of OTE.

Bacterial Adhesion↗

PubMed1993

Unexpectedly high levels of some presumably protective secretory immunoglobulin A antibodies to dental plaque bacteria in salivas of both caries-resistant and caries-susceptible subjects.

The role of salivary antibodies in protection against cariogenic bacteria is actually a matter of debate. Correlation between caries experience and naturally occurring antibodies was extensively investigated. Comparison of salivary antibodies from 21 caries-resistant and 22 caries-susceptible subjects was carried out by using a new quantitative method. Secretory immunoglobulin A (S-IgA) antibodies to Streptococcus sobrinus and Streptococcus sanguis cells were detected in all salivas and at similar levels in both groups. When assayed with two major antigens from S. sobrinus, i.e., protein antigen I/II and cell wall carbohydrates, only specific activities of antibodies to the protein component were increased (P < 0.01), but this occurred unexpectedly in the caries-susceptible group. Western blot (immunoblot) analysis with the culture supernatant and cell wall proteins from S. sobrinus showed the same antibody specificity in both groups. No selective increase of the protease-resistant S-IgA2 subclass was found, and avidities of antibodies to both antigen I/II and cell wall carbohydrates were similar. Our results demonstrate that naturally induced S-IgA antibodies against S. sanguis, S. sobrinus, and the major antigens of the latter are not sufficient to inhibit caries development.

Adult↗

PubMed1993

Antigenicity and immunogenicity of a synthetic peptide derived from a glucan-binding domain of mutans streptococcal glucosyltransferase.

The immunogenicity and antigenicity of a multiply antigenic peptide construct containing four copies of the synthetic peptide TGAQTIKGQKLYFKANGQQVKG were measured in rodents and humans, respectively. The composition of this peptide construct (termed GLU) was derived from a major repeating sequence in the C-terminal region of mutans streptococcal glucosyltransferases that synthesize water-insoluble glucan (GTF-I). The GLU peptide elicited high levels of serum immunoglobulin G antibody to GLU after subcutaneous injection into Sprague-Dawley rats. These antisera also reacted with intact GTF isozymes from Streptococcus sobrinus and Streptococcus mutans (by enzyme-linked immunosorbent assay [ELISA] and Western blot [immunoblot] analyses) and with an 87-kDa glucan-binding protein from S. sobrinus (by Western blot). The synthesis of filter-retained glucan by GTF-Sd of S. sobrinus could be inhibited (30%) by preincubation with anti-GLU rat serum. Splenic and lymph node lymphocytes from rats injected once with S. sobrinus GTF isozymes demonstrated significant proliferation after 5 days of culture with GLU. The GLU peptide reacted with 4 of 29 human parotid saliva samples and 5 of 29 human serum samples (by ELISA). These results suggest that the GLU peptide contains B- and T-cell epitopes that are similar to those of intact mutans streptococcal GTFs and possibly certain other glucan-binding proteins as well. Furthermore, since antibody to this epitope(s) appears to inhibit GTF function, sequences within this peptide construct may have value for inclusion in a synthetic dental caries vaccine.

Adult↗